Molecular Oncology Group, Biodonostia Research Institute, 20014 San Sebastián, Spain.
Sino-Swiss Institute of Advanced Technology (SSIAT), Shanghai University, Shanghai 201907, China.
Biosensors (Basel). 2023 Feb 15;13(2):275. doi: 10.3390/bios13020275.
Even with the widespread uptake of vaccines, the SARS-CoV-2-induced COVID-19 pandemic continues to overwhelm many healthcare systems worldwide. Consequently, massive scale molecular diagnostic testing remains a key strategy to control the ongoing pandemic, and the need for instrument-free, economic and easy-to-use molecular diagnostic alternatives to PCR remains a goal of many healthcare providers, including WHO. We developed a test (Repvit) based on gold nanoparticles that can detect SARS-CoV-2 RNA directly from nasopharyngeal swab or saliva samples with a limit of detection (LOD) of 2.1 × 10 copies mL by the naked eye (or 8 × 10 copies mL by spectrophotometer) in less than 20 min, without the need for any instrumentation, and with a manufacturing price of <$1. We tested this technology on 1143 clinical samples from RNA extracted from nasopharyngeal swabs ( = 188), directly from saliva samples ( = 635; assayed by spectrophotometer) and nasopharyngeal swabs ( = 320) from multiple centers and obtained sensitivity values of 92.86%, 93.75% and 94.57% and specificities of 93.22%, 97.96% and 94.76%, respectively. To our knowledge, this is the first description of a colloidal nanoparticle assay that allows for rapid nucleic acid detection at clinically relevant sensitivity without the need for external instrumentation that could be used in resource-limited settings or for self-testing.
即使广泛采用疫苗，SARS-CoV-2 引起的 COVID-19 大流行仍继续使全球许多医疗系统不堪重负。因此，大规模分子诊断检测仍然是控制当前大流行的关键策略，包括世界卫生组织在内的许多医疗保健提供者仍需要无仪器、经济且易于使用的分子诊断替代 PCR。我们开发了一种基于金纳米粒子的检测方法（Repvit），可以直接从鼻咽拭子或唾液样本中检测 SARS-CoV-2 RNA，通过肉眼检测的检出限（LOD）为 2.1×10 拷贝/mL（通过分光光度计检测为 8×10 拷贝/mL），检测时间不到 20 分钟，无需任何仪器，且制造成本<$1。我们在来自多个中心的 1143 份鼻咽拭子 RNA 提取样本（ = 188）、直接来自唾液样本（ = 635；通过分光光度计检测）和鼻咽拭子样本（ = 320）上测试了这项技术，获得了 92.86%、93.75%和 94.57%的敏感性值，以及 93.22%、97.96%和 94.76%的特异性值。据我们所知，这是第一个描述胶体纳米粒子检测方法的报道，该方法可在临床相关的灵敏度下无需外部仪器进行快速核酸检测，适用于资源有限的环境或用于自我检测。
